The present disclosure relates to cell-free compositions and methods for making and using the same. In one aspect, the composition includes: an extract derived from one or more organisms; one or more proteins of interest, wherein the one or more proteins are expressed from one or more nucleic acids exogenous to the extract and/or by the one or more organisms, wherein preferably the one or more proteins react with a substrate to produce a product; and one or more O2, O—, or H.sub.2O.sub.2 scavengers. The composition may be oxygen-deprived. The composition may also include an energy recycling system.

The present disclosure relates to genetically engineered methanotrophic bacteria with the capability of growing on a multi-carbon substrate as a primary or sole carbon source and methods for growing methanotrophic bacteria on a multi-carbon substrate.

The present invention relates to the field of genetic engineering, particularly to a glucose oxidase mutant having improved thermal stability, gene and application thereof. The present invention provides several glucose oxidase GOD mutants with high catalytic efficiency and improved thermal stability, which breaks the barrier of low enzyme activity and poor stability and is suited well to meet the requirements of application to the fields of food, medicine, feed and textile industry, and has a very broad application prospect.

The present disclosure relates to a multi-enzyme conjugate, a method for preparing the same and a method for preparing an organic compound using the same. More particularly, a multi-enzyme conjugate exhibiting improved catalytic efficiency over respective free enzymes using site-specific incorporation of a clickable non-natural amino acid into the enzymes and two compatible click reactions, a method for preparing the same and a method for preparing an organic compound using the same may be provided.

The present invention relates to: a novel Pichia kudriavzevii microorganism NG7 showing heat resistance and acid resistance; a composition, for producing organic acid or alcohol, which comprises the microorganism and a culture of the same; and a method, for producing an organic acid or alcohol, which comprises culturing the microorganism.

An object of the present invention is to obtain a fermentative yeast having a highly efficient ethanol production without introducing a foreign gene. A further object is to obtain a fermentative yeast that is resistant to proliferation inhibitors such as organic acids, which prevent the growth of the fermentative yeast. Yeast having improved ethanol production ability was generated by introducing transaldolase and alcohol dehydrogenase gene by self-cloning to Meyerozyma guilliermondii that can produce ethanol effectively from pentose and hexose obtained by breeding. This fermentative yeast is deposited to NITE Patent Microorganisms Depositary under the accession number NITE ABP-01976.

Disclosed is a microorganism, which has been engineered to acquire methylotrophy. More particularly, the application describes a non-naturally occurring microorganism, which has been engineered to express or include a first enzyme and a second enzyme, wherein the first enzyme is a methanol dehydrogenase (Mdh) enzyme or a methanol oxidase (Mox) enzyme, and wherein the second enzyme is a dihydroxyacetone synthase (Das) enzyme or a transketolase enzyme. The application also describes element and applications, more particularly kits and methods, which include or use the microorganism.

The present disclosure provides methods and compositions for producing rotundone. In various aspects, the present disclosure provides enzymes, polynucleotides encoding said enzymes, and recombinant microbial host cells (or microbial host strains) for the production of rotundone. In some embodiments, the present disclosure provides microbial host cells for producing rotundone at high purity and/or yield, from either enzymatic transformation of α-guaiene, or from sugar or other carbon source. The present disclosure further provides methods of making products containing rotundone, including flavor or fragrance products, among others.

A method of producing a modified Saccharomyces cerevisiae yeast strain with enhanced resistance (or tolerance) to pretreatment-derived microbial inhibitors such as furans, phenolics and weak acids is provided, which comprises integrating at least one copy of the TAL1 gene and at least one copy of two or more of the FDH1, AR11 and ADH6 genes into the S. cerevisiae genome. A modified yeast strain so obtained is also provided, the modified yeast strain being capable of simultaneously overexpressing these genes relative to a yeast strain which hasn't been modified in the same manner. S. cerevisiae strains which have been modified as described herein can be used to ferment lignocellulosic hydrolysates containing pretreatment inhibitors such as furans, phenolics and weak acids. Suitable lignocellulosic hydrolysates include sugarcane bagasse (SCB) and waste streams from the pulp and paper industry, such as spent sulphite liquor (SSL).

Recombinant cells and methods for producing methyl ketones, such as medium-chain methyl ketones. The recombinant cells include recombinant acyl-ACP thioesterase genes, recombinant β-ketoacyl-CoA thioesterase genes, and recombinant acyl-CoA synthetase genes, in addition to other modifications. The methods include culturing the recombinant cells to produce the methyl ketones and isolating the produced methyl ketones.